Comprehensive two-dimensional gas chromatography–time-of-flight mass spectrometry (GC × GC-TOF) for high resolution metabolomics: biomarker discovery on spleen tissue extracts of obese NZO compared to lean C57BL/6 mice

Abstract

Comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (GC · GC-TOF) was applied for the analysis of complex metabolite profiles from mouse spleen. The resulting two-dimensional chromatograms proved that mass spectral quality and sensitivity were largely improved by the enhanced resolution and zone compression, which are features of GC · GC operation, when compared to classical one-dimensional GC-TOF methods. The improved peak capacity of GC · GC allowed for peaks to be detected that could previously not be separated in one-dimensional GC. A measure of the combined power of chromatographic and mass spectral deconvolution resolution is called ''analytical purity'', with higher values indicating less pure peaks. GC · GC-TOF lead to the detection of 1200 compounds with purity better than 0.2, compared to 500 compounds with purity up to 2.5 in one-dimensional GC-TOF. The compounds identified include many of the compounds previously reported in NMR studies and other methods on mammalian tissues. Spleen samples of several obese NZO mice and lean C57BL/6 control strains were analyzed in order to demonstrate the applicability of GC · GC-TOF for biomarker identification.

Comprehensive two‐dimensional gas chromatography‐mass spectrometry: A review

“…In the 2004-2005 period, there was a significant interest towards the analysis of metabolites by using GC Â GC-ToF-MS (Sinha et al, 2004b,c;Hope et al, 2005b;Shellie et al, 2005;Welthagen et al, 2005). The analytical method appeared to be particularly suited, because the determination of volatile metabolites in a specific biological situation is a large task.…”

“…The analytical method appeared to be particularly suited, because the determination of volatile metabolites in a specific biological situation is a large task. Welthagen et al (2005) applied GC and GC Â GC, both combined with ToF-MS, to the analysis of spleen extracts of obese (NZO strain) and lean (C57BL/6 strain) mice. A 10-fold higher sample amount was injected onto the single GC column to compensate for the lower sensitivity.…”

Mass Spectrometry–Based Metabolic Profiling Reveals Different Metabolite Patterns in Invasive Ovarian Carcinomas and Ovarian Borderline Tumors

“…Metabolite profiling has been used in clinical research to study metabolic diseases (8,9); however, surprisingly few reports (10,11) have studied comprehensive metabolic responses for the characterization of tumor types. The combination of gas chromatography-TOF (GC-TOF)-based analysis with automatical deconvolution techniques has not been used for analysis of human tumor samples, thus far, but only on mouse tissues in obesity-related research (12).…”

The Interplay of Proton, Electron, and Metabolite Supply for Photosynthetic H2 Production in Chlamydomonas reinhardtii

“…This article reports on the differential analysis of metabolite profiles between Stm6Glc4, the newest and most advanced generation of our high H 2 -producing C. reinhardtii strains (18), and the corresponding WT cc406 before and during H 2 production using two-dimensional GC coupled with time of flight MS (GCxGC-TOFMS), which has resolved an order of magnitude more compounds than previously reported (29). Stm6Glc4 is a cell line developed by inserting a hexose symporter into the high H 2 -producing mutant Stm6.…”